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Drawing method of acute hepatic failure mouse liver full-immune cell characteristic spectrum

A technology of acute liver failure and immune cells, which is applied in the field of drawing the characteristic map of liver immune cells in mice with acute liver failure, can solve the problems of insufficient coverage of liver immune cells and inability to accurately reflect the characteristic changes of liver immune cell subsets

Inactive Publication Date: 2019-10-15
ZHEJIANG UNIV
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Problems solved by technology

Due to technical limitations such as limited detection channels and overlapping emission spectra of fluorophores in traditional flow cytometry, experiments with more than 12 colors cannot be carried out, resulting in insufficient coverage of liver immune cell detection, and cannot accurately reflect the subset of liver immune cells in liver failure. group characteristic change

Method used

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  • Drawing method of acute hepatic failure mouse liver full-immune cell characteristic spectrum
  • Drawing method of acute hepatic failure mouse liver full-immune cell characteristic spectrum
  • Drawing method of acute hepatic failure mouse liver full-immune cell characteristic spectrum

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Embodiment Construction

[0049] At first the source of the mouse liver used in the present invention is described:

[0050] The mouse livers used in the present invention are all taken from the discarded mice in the laboratory that died naturally due to acute liver failure, and the removal of the liver should be within 1 hour of the death time. Among them, the corpses of C57BL / 6J young mice aged 6-8 weeks and weighing 18-25 g were selected, washed with 100 ml of 70% ethanol, and the liver was taken out in a biological safety cabinet for subsequent tests. During the implementation of the present invention, there is no operation of killing surviving mice, and there is no traumatic or interventional treatment such as dissection and excision of living mice.

[0051] The technical solutions of the present invention will be described in detail below in conjunction with specific implementation examples.

[0052] Instruments and Reagents

[0053] Dissociation tube (MACS, German), dissociation machine (MACS,...

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Abstract

The invention relates to the liver full-immune cell characteristic spectrum research, and aims at provided a drawing method of acute hepatic failure mouse liver full-immune cell characteristic spectrum. The method comprises the following steps: mouse liver full-immune cells extraction; mass spectrum flow antibody marking; immune cell antibody dyeing; and mass spectrum flow cytometry. On the basisof guaranteeing the cell purity, the liver parenchyma cell can be removed as much as possible, thereby separating out the mouse liver full-immune cells in high yield. The yield of the separated mouseliver full-immune cell is higher than the yield of the grinding method, and the cell survival rate is greater than that of the traditional grinding method. According to immune cell features in the liver and lung, a kit is innovatively used for stabilizing the metal isotope and antibody; a mass spectrum flow channel is designed based on the channel minimum interference principle, the classificationand function of the liver full-immune cell can be comprehensively described, at most 43 markers can be detected at the same time, and the dynamic change of the mouse liver full-immune cell can be observed.

Description

[0001] field of invention [0002] The invention relates to the research on the characteristic atlas of whole immune cells in the liver, in particular to a method for drawing the characteristic atlas of the whole immune cells in the liver of mice with acute liver failure. Background technique [0003] Acute liver failure is a group of severe clinical syndromes in which a large number of hepatic cell necrosis and severe liver function damage occur within a short period of time, leading to hepatic encephalopathy. The mortality rate of this disease is high, and liver transplantation is the exact and effective treatment at present. However, factors such as organ shortage and post-transplantation complications greatly limit the clinical application of liver transplantation. In recent years, immune agents, stem cell transplantation and other treatments aimed at pathogenesis have developed rapidly and are expected to become new treatments. [0004] During liver failure, a large num...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/60G01N33/532G01N27/62G01N15/14G01N1/28G01N33/50
CPCG01N33/6848G01N33/6854G01N33/60G01N33/532G01N15/14G01N27/626G01N1/28G01N33/5005G01N2560/00
Inventor 曹红翠刘景琪冯旭东俞炯潘巧玲徐燕萍曾浔李兰娟
Owner ZHEJIANG UNIV
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